This invention relates to aminobutyric acid derivatives, processes for the preparation thereof and pharmaceutical agents containing them as active ingredient. More particularly, this invention relates to:
aminobutyric acid derivatives of the formula (I): 
wherein all the symbols are as hereinafter defined, and non-toxic salts thereof, processes for the preparation thereof and pharmaceutical agents containing them as active ingredient.
The matrix metalloproteinases (MMPS) are neutral metalloproteinases and zinc (Zn2+) is essential in the active site for their activation. They degrade collagen, laminin, proteoglycans, fibronectin, elastin, gelatin etc. under physiological conditions and therefore, are effective on growth and tissue remodeling of articulation tissue, bone tissue and connective tissue. At least 10 classes of MMPs which differ in primary structure are identified. Concretely, there are Interstitial Coliagenase (MMP-1), Neutrophil Collagenase (MMP-8), Gelatinase A (MMP-2), Gelatinase B (MMP-9), Stromelysin-1 (MMP-3), Stromelysin-2 (MMP-10), Matrilysin (MMP-7), metalloerastase (MMP-12) etc.
As common characteristics of these enzymes, MMPs
(1) have Zn2+ in the active site and the activity depends on calcium ion (Ca2+),
(2) are secreted as an inactive proenzyme and activated outside of cells,
(3) have high homology on amino acid sequence,
(4) have an ability to degrade on various extracellular matrix components in vivo,
(5) are regulated by tissue inhibitors of metalloproteinases (TIMP) which are specific to MMPs.
MMP inhibitors are useful for prevention and/or treatment of various diseases induced by overexpression and excess activation of MMP. Such diseases are, for example, rheumatoid disease, arthrosteitis, unusual bone resorption, osteoporosis, periodontitis, interstitial nephritis, arteriosclerosis, pulmonary emphysema, cirrhosis, cornea injury, metastasis of, invasion of or growth of tumor cells, autoimmune diseases (e.g. Crohn""s disease, Sjogren""s syndrome), diseases caused by vascular emigration or infiltration of leukocytes, arterialization, multiple sclerosis, aorta aneurysm, endometriosis.
Some compounds possessing inhibitory activity against MMP are known. A sequence in the vicinity of cleavage site of collagen (Gly-Ile-Ala-Gly or Gly-Leu-Ala-Gly) has high affinity for collagenase.
Much research and development on substrate analogous MMP inhibitors, which are chemically modified so as to have zinc affinity groups on a cleaving site of the substrate, has energetically been carried out [Inhibitors of matrix metalloproteinases (MMP""s), Nigel R A Beeley, Phillip R J Ansell, Andrew J P Docherty et al., Curr. Opin. Ther. Patents., 4, 7-16 (1994), Current Drugs Ltd ISSN 0962-2594]. However, these substrate-analogues inhibitors might have various problems. Therefore, it is desired to obtain a non-peptide inhibitor and some compounds are reported.
For example, in the specification of EP 757037 as the Example, sulfonylamino acid derivatives of the formula (W): 
are disclosed to have an activity of inhibiting matrix metalloproteinase.
In the specification of EP 757984 as the Example, hydroxamic acid derivatives of the formula (X): 
are disclosed to have an activity of inhibiting matrix metalloproteinase.
In the specification of WO 9723459 as the Example, aromatic keto-acid derivatives of the formula (Y): 
are disclosed to have an activity of inhibiting matrix metalloproteinase.
In the specification of WO 9718188 as the Example, hydroxamic acid derivatives of the formula (Z): 
are disclosed to have an activity of inhibiting matrix metalloproteinase and TNFxcex1 secretion.
Energetic investigations have been carried out in order to make a matrix metalloproteinase, e.g. gelatinase, stromelysin or collagenase, inhibitor. The present inventors have found that novel compounds of aminobutyric acid derivatives of the formula (I) which are carboxylic amino derivatives of xcex3-amino acid, accomplished the present purpose.
The present invention relates to:
1) an aminobutyric acid derivative of the formula (I): 
xe2x80x83wherein
R1 is xe2x80x94COOR10, xe2x80x94CONHOR10, xe2x80x94CONHNHR10, xe2x80x94(CH2)nSR50 or xe2x80x94Yxe2x80x94P(OR51)2;
R10 is (i) hydrogen, (ii) C1-8 alkyl, (iii) phenyl, (iv) C1-8 alkyl substituted by phenyl or C1-8 alkoxy, or (v) oxycarbonyl substituted by phenyl, benzyl or C1-8 alkyl;
n is 0-3;
R50 is (i) hydrogen, (ii) C1-8 alkyl, (iii) xe2x80x94COR52, in which R52 C1-8 alkyl or phenyl; or (iv) xe2x80x94SR53, in which R53 is hydrogen, C1-8 alkyl or phenyl;
R51 is hydrogen, C1-8 alkyl or phenyl;
Y is a single bond, xe2x80x94CH2xe2x80x94 or xe2x80x94Oxe2x80x94;
R2, R3, R4, R5, R6 and R7 each, independently, is
(1) hydrogen,
(2) C1-8 alkyl,
(3) C2-8 alkenyl,
(4) xe2x80x94OR11,
(5) xe2x80x94SR11 ,
(6) xe2x80x94NR12R13,
(7) xe2x80x94COR14,
(8) Cyc1,
(9) C1-8 alkyl substituted by xe2x80x94OR11, xe2x80x94SR11, xe2x80x94NR12R13, xe2x80x94COR14, guanidino or Cyc1, or
(10) C2-8 alkenyl substituted by xe2x80x94OR11, xe2x80x94SR11, xe2x80x94NR12R13, xe2x80x94COR14, guanidino or Cyc1, or
R2 and R4, taken together is C1-8 alkylene, R5 and R6, taken together is C1-8 alkylene, R3 and R6, taken together is C1-8 alkylene, R2 and R3, taken together is C2-8 alkylene, R4 and R5, taken together is C2-8 alkylene, or R6 and R7, taken together is C2-8 alkylene;
in which Cyc1 is carbocyclic ring or heterocyclic ring and these carbocyclic ring and heterocyclic ring may be substituted by one or more of (i) C1-8 alkyl, (ii) C1-8 alkoxy, (iii) nitro, (iv) guanidino, (v) amidino, (vi) halogen atom, (vii) nitrile (viii) hydroxy, (ix) benzyloxy, (x) xe2x80x94NR101R102, in which R101 and R102 each, independently, is hydrogen or C1-8 alkyl, (xi) xe2x80x94COOR103, in which R103 is hydrogen or C1-8 alkyl, (xii) trifluoromethyl, (xiii) trifluoromethyloxy, (xiv) phenyl, (xv) phenyl substituted by C1-8 alkyl or C1-8 alkoxy, (xvi) phenyloxy, (xvii) phenylsulfonyl, (xviii) C1-8 alkyl substituted by phenyl or nitrile, (xix) heterocyclic ring, (xx) keto, and (xxi) C1-8 alkoxy substituted by xe2x80x94CONR104R105, in which R104 and R105 each, independently, is hydrogen, C1-8 alkyl or phenyl;
R11 is (i) hydrogen, (ii) C1-8 alkyl, (iii) Cyc1, or (iv) xe2x80x94COR18, or C1-8 alkyl substituted by xe2x80x94OR15, xe2x80x94SR15, xe2x80x94NR16R17, xe2x80x94COR18, guanidino or Cyc1;
R15 is hydrogen, C1-8 alkyl, Cyc1 or C1-8 alkyl substituted by Cyc1 or C1-8 alkoxy;
R16 is hydrogen or C1-8 alkyl;
R17 is hydrogen, C1-8 alkyl or xe2x80x94COR19, in which R19 is C1-8 alkyl, Cyc1 or C1-8 alkyl substituted by Cyc1;
R18 is hydroxy, C1-8 alkyl, C1-8 alkoxy or xe2x80x94NR20R21, in which R20 and R21, each independently, is hydrogen, C1-8 alkyl, Cyc1 or C1-8 alkyl substituted by Cyc1;
R12 is hydrogen, C1-8 alkyl, Cyc1 or C1-8 alkyl substituted by Cyc1;
R13 is hydrogen, C1-8 alkyl, Cyc1, C1-8 alkyl substituted by Cyc1, or xe2x80x94COR22, in which R22 is C1-8 alkyl, Cyc1 or C1-8 alkyl substituted by Cyc1;
R14 is hydroxy, C1-8 alkyl, C1-8 alkoxy, Cyc1, C1-8 alkyl substituted by Cyc1, or xe2x80x94NR23R24, in which R23 and R24, each independently, is (i) hydrogen, (ii) C1-8 alkyl, (iii) Cyc1 or (iv) C1-8 alkyl substituted by Cyc1 or hydroxy;
(1) R8 is
1) hydrogen,
2) C1-8 alkyl,
3) C1-8 alkoxycarbonyl,
4) C1-8 alkyl substituted by xe2x80x94OR26, xe2x80x94SR26, xe2x80x94NR27R28 or xe2x80x94COR29, or
5) C1-8 alkoxycarbonyl substituted by Cyc2, and
R9 is 
(2) R8 is 
R9 is
1) C1-8 alkyl,
2) C1-8 alkoxy,
3) C1-8 alkoxy substituted by Cyc2,
4) C1-8 alkyl substituted by xe2x80x94OR26, xe2x80x94SR26, xe2x80x94NR27R28, xe2x80x94COR29 or Cyc2 or
5) 
xe2x80x83in which Cyc2 is carbocyclic ring or heterocyclic ring and these carbocyclic ring and heterocyclic ring may be substituted by one or more of (i) C1-8 alkyl, (ii) C1-8 alkoxy, (iii) nitro, (iv) guanidino, (v) amidino, (vi) halogen atom, (vii) nitrile (viii) hydroxy, (ix) benzyloxy, (x) xe2x80x94NR201R202, in which R201 and R202 each, independently, is hydrogen or C1-8 alkyl, (xi) xe2x80x94COOR203, in which R203 is hydrogen or C1-8 alkyl, (xii) trifluoromethyl, (xiii) trifluoromethyloxy, (xiv) phenyl, (xv) phenyl substituted by C1-8 alkyl or C1-8 alkoxy, (xvi) phenyloxy, (xvii) phenylsulfonyl, (xviii) C1-8 alkyl substituted by phenyl or nitrite, (xix) heterocyclic ring, (xx) keto, and (xxi) C1-8 alkoxy substituted by xe2x80x94CONR204R205, in which R204 and R205 each, independently, is hydrogen, C1-8 alkyl or phenyl;
R26 is hydrogen, C1-8 alkyl, Cyc2 or C1-8 alkyl substituted by Cyc2;
R27 is hydrogen, C1-8 alkyl, Cyc2 or C1-8 alkyl substituted by Cyc2;
R28 is hydrogen, C1-8 alkyl, Cyc2, C1-8 alkyl substituted by Cyc2, or xe2x80x94COR30, in which R30 is C1-8 alkyl, Cyc2 or C1-8 alkyl substituted by Cyc2;
R29 is hydroxy, C1-8 alkyl, Cyc2, C1-8 alkyl substituted by Cyc2, or xe2x80x94NR31R32, in which R31 and R32, each independently, is hydrogen, C1-8 alkyl, Cyc2 or C1-8 alkyl substituted by Cyc2; 
xe2x80x83is carbocyclic ring or heterocyclic ring;
R25 is xe2x80x94Exe2x80x94G;
E is
1) a single bond,
2) xe2x80x94CONR33xe2x80x94,
3) xe2x80x94NR33COxe2x80x94,
4) xe2x80x94COxe2x80x94Oxe2x80x94,
5) xe2x80x94Oxe2x80x94COxe2x80x94,
6) xe2x80x94NR33xe2x80x94COxe2x80x94NR34xe2x80x94,
7) xe2x80x94COxe2x80x94CH2xe2x80x94,
8) xe2x80x94COxe2x80x94,
9) xe2x80x94Oxe2x80x94COxe2x80x94NR33xe2x80x94,
10) xe2x80x94NR33xe2x80x94COxe2x80x94Oxe2x80x94,
11) xe2x80x94Oxe2x80x94COxe2x80x94Oxe2x80x94,
12) xe2x80x94CSxe2x80x94NR33xe2x80x94,
13) xe2x80x94NR33xe2x80x94CSxe2x80x94,
14) xe2x80x94CSxe2x80x94Oxe2x80x94,
15) xe2x80x94Oxe2x80x94CSxe2x80x94,
16) xe2x80x94NR33xe2x80x94CSxe2x80x94R34xe2x80x94,
17) xe2x80x94CSxe2x80x94CH2xe2x80x94,
18) xe2x80x94CSxe2x80x94,
19) xe2x80x94Oxe2x80x94CSxe2x80x94NR33xe2x80x94,
20) NR33xe2x80x94CSxe2x80x94Oxe2x80x94,
21) xe2x80x94Oxe2x80x94CSxe2x80x94Oxe2x80x94,
22) xe2x80x94CH2xe2x80x94CH2xe2x80x94,
23) xe2x80x94HCxe2x95x90CHxe2x80x94,
24) xe2x80x94Cxe2x89xa1Cxe2x80x94,
25) xe2x80x94SO2xe2x80x94NR33xe2x80x94,
26) xe2x80x94NR33xe2x80x94SO2xe2x80x94,
27) xe2x80x94SO2xe2x80x94CH2xe2x80x94 or
28) xe2x80x94CH2xe2x80x94SO2xe2x80x94;
R33 and R34, each independently, is hydrogen, C1-8 alkyl, Cyc3 or C1-8 alkyl substituted by Cyc3;
Cyc 3 is carbocyclic ring or heterocyclic ring and these carbocyclic ring and heterocyclic ring may be substituted by one or more of (i) C1-8 alkyl, (ii) C1-8 alkoxy, (iii) nitro, (iv) guanidino, (v) amidino, (vi) halogen atom, (vii) nitrile, (viii) hydroxy, (ix) benzyloxy, (x) xe2x80x94NR301R302, in which R301 and R302 each, independently, is hydrogen or C1-8 alkyl, (xi) xe2x80x94COOR303, in which R303 is hydrogen or C1-8 alkyl, (xii) trifluoromethyl, (xiii) trifluoromethyloxy, (xiv) phenyl, (xv) phenyl substituted by C1-8 alkyl or C1-8 alkoxy, (xvi) phenyloxy, (xvii) phenylsulfonyl, (xviii) C1-8 alkyl substituted by phenyl or nitrite, (xix) heterocyclic ring, (xx) keto, and (xxi) C1-8 alkoxy substituted by xe2x80x94CONR304R305, in which R304 and R305 each, independently, is hydrogen, C1-8 alkyl or phenyl;
G is
1) hydrogen,
2) C1-8 alkyl,
3) Cyc4,
4) xe2x80x94OR35,
5) xe2x80x94SR35,
6) halogen atom,
7) nitro,
8) nitrite,
9) xe2x80x94NR36R37,
10) xe2x80x94COR38,
11) C1-8 alkyl substituted by Cyc4, xe2x80x94OR35, xe2x80x94SR35, halogen atom, xe2x80x94NR36R37 or xe2x80x94COR38;
in which Cyc4 is carbocyclic ring or heterocyclic ring and these carbocyclic ring and heterocyclic ring may be substituted by one or more of (i) C1-8 alkyl, (ii) C1-8 alkoxy, (iii) nitro, (iv) guanidino, (v) amidino, (vi) halogen atom, (vii) nitrile (viii) hydroxy, (ix) benzyloxy, (x) xe2x80x94NR401R402, in which R401 and R402 each, independently, is hydrogen or C1-8 alkyl, (xi) xe2x80x94COOR403, in which R403 is hydrogen or C1-8 alkyl, (xii) trifluoromethyl, (xiii) trifluoromethyloxy, (xiv) phenyl, (xv) phenyl substituted by C1-8 alkyl or C1-8 alkoxy, (xvi) phenyloxy, (xvii) phenylsulfonyl, (xviii) C1-8 alkyl substituted by phenyl or nitrile, (xix) heterocyclic ring, (xx) keto, and (xxi) C1-8 alkoxy substituted by xe2x80x94CONR404R405, in which R404 and R405 each, independently, is hydrogen, C1-8 alkyl or phenyl;
R35 is hydrogen, C1-8 alkyl, C1-8 alkoxy, Cyc4 or C1-8 alkyl substituted by Cyc4;,
R36 is hydrogen, C1-8 alkyl, Cyc4 or C1-8 alkyl substituted by Cyc4;
R37 is hydrogen, C1-8 alkyl, Cyc4, C1-8 alkyl substituted by Cyc4, or xe2x80x94COR39, in which R39 is C1-8 alkyl, Cyc4 or C1-8 alkyl substituted by Cyc4;
R38 is hydroxy, C1-8 alkyl, Cyc4, C1-8 alkyl substituted by Cyc4, or xe2x80x94NR40R41, in which R40 and R41, each independently, is hydrogen, C1-8 alkyl, Cyc4 or C1-8 alkyl substituted by Cyc4; or
xe2x80x94Exe2x80x94G taken together, is C1-4 alkylidene;
p is 1-5;
M is C1-8 alkylene;
J is a single bond, an oxygen atom, a sulfur atom or xe2x80x94NR42xe2x80x94, in which R42 is hydrogen or C1-8 alkyl;
 may be double bond, by releasing the hydrogens, when two of R2, R3, R4, R5, R6 and R7 that do not bond to the same carbon atom and bond to neighboring carbon atoms, are hydrogens, with the proviso that  is not double bond, when R3 and R4, taken together is C1-8 alkylene, R5 and R6, taken together is C1-8 alkylene, or R3 and R6, taken together is C1-8 alkylene;
or a non-toxic salt thereof,
2) a process for the preparation of an aminobutyric acid derivative of the formula (I) or a non-toxic salt thereof, and
3) a pharmaceutical agent containing an aminobutyric acid derivative of the formula (I) or a non-toxic salt thereof as active ingredient.
Unless otherwise specified, all isomers are included in the present invention. For example, alkyl, alkoxy and alkylene include straight and branched isomers. Double bond in alkenylene includes structure of configurations E, Z and EZ mixture. Isomers resulting from the presence of asymmetric carbon(s) e.g. branched alkyl, alkoxy and alkylene are also included within the present invention.
In the present invention, C1-8 alkyl is methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl and isomeric groups thereof.
C1-8 alkoxy is methoxy, ethoxy, propoxy, butoxy, pentyloxy, hexyloxy, heptyloxy, octyloxy and isomeric groups thereof.
C1-8 alkyl substituted by phenyl is methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl and isomeric groups thereof substituted by one of phenyl.
C1-8 alkyl substituted by C1-8 alkoxy is methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl and isomeric groups thereof substituted by one of methoxy, ethoxy, propoxy, butoxy, pentyloxy, hexyloxy, heptyloxy, octyloxy and isomeric groups thereof.
C1-8 alkyl substituted by nitrile is methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl and isomeric groups thereof substituted by one of nitrile.
Oxycarbonyl substituted by phenyl is phenyloxycarbonyl.
Oxycarbonyl substituted by benzyl is benzyloxycarbonyl.
Oxycarbonyl substituted by C1-8 alkyl is methyloxycarbonyl, ethyloxycarbonyl, propyloxycarbonyl, butyloxycarbonyl, pentyloxycarbonyl, hexyloxycarbonyl, heptyloxycarbonyl, octyloxycarbonyl and isomeric groups thereof.
C2-8 alkenyl is vinyl, propenyl, butenyl, pentenyl, hexenyl, heptenyl, octenyl, butadienyl, pentadienyl, hexadienyl, heptadienyl, octadienyl, hexatrienyl, heptatrienyl, octatrienyl and isomeric groups thereof.
C1-8 alkylene is methylene, ethylene, trimethylene, tetramethylene, pentamethylene, hexamethylene, heptamethylene, octamethylene, and isomeric groups thereof.
C2-8 alkylene is ethylene, trimethylene, tetramethylene, pentamethylene, hexamethylene, heptamethylene, octamethylene, and isomeric groups thereof.
Halogen atom is chlorine, bromine, fluorine, or iodine.
C1-8 alkoxycarbonyl is methyloxycarbonyl, ethyloxycarbonyl, propyloxycarbonyl, butyloxycarbonyl, pentyloxycarbonyl, hexyloxycarbonyl, heptyloxycarbonyl, octyloxycarbonyl and isomeric groups thereof.
C1-4 alkylidene is methylidene, ethylidene, propylidene, butylidene and isomeric groups thereof.
Carbocyclic ring is C3-15 mono-, bi- or tri-carbocyclic ring, for example, cyclopropane, cyclobutane, cyclopentane, cyclohexane, cycloheptane, cyclopentene, cyclohexene, cyclopentadiene, cyclohexadiene, benzene, pentalene, indene, naphthalene, azulene, fluorene, phenanthrene, anthracene, acenaphthylene, biphenylene, perhydropentalene, perhydroindene, perhydronaphthalene, perhydroazulene, perhydrofluorene, perhydrophenanthrene, perhydroanthracene, perhydroacenaphthylene, perhydrobiphenylene, adamantane.
Heterocyclic ring is 5-18 membered mono-, bi- or tri-heterocyclic ring containing 1-4 of nitrogen(s), 1-2 of oxygen(s) and/or 1-2 of sulfur(s). 5-18 membered mono-, bi- or tri-heterocyclic ring containing 1-4 of nitrogen(s), 1-2 of oxygen(s) and/or 1-2 of sulfur(s) includes 5-18 membered mono-, bi- or tri-heterocyclic aryl containing 1-4 of nitrogen(s), 1-2 of oxygen(s) and/or 1-2 of sulfur(s), and partially or fully saturated thereof.
5-18 membered mono-, bi- or tri-heterocyclic aryl containing 1-4 of nitrogen(s), 1-2 of oxygen(s) and/or 1-2 of sulfur(s), is, for example, pyrrole, imidazole, triazole, tetrazole, pyrazole, pyridine, pyrazine, pyrimidine, pyridazine, azepine, diazepine, furan, pyran, oxepin, oxazepine, thiophene, thiain (thiopyran), thiepin, oxazole, isoxazole, thiazole, isothiazole, oxadiazole, oxazine, oxadiazine, oxazepine, oxadiazepine, thiadiazole, thiazine, thiadiazine, thiazepine, thiadiazepine, indole, isoindole, benzofuran, isobenzofuran, benzothiophene, isobenzothiophene, indazole, quinoline, isoquinoline, phthalazine, naphthyridine, quinoxaline, quinazoline, cinnoline, benzoxazole, benzothiazole, benzoimidazole, carbazole or acridine.
Partially or fully saturated 5-18 membered mono- or bi-heterocyclic ring containing 1-4 of nitrogen(s), 1-2 of oxygen(s) and/or 1-2 of sulfur(s), for example, pyrroline, pyrrolidine, imidazoline, imidazolidine, triazoline, triazolidine, tetrazoline, tetrazolidine, pyrazoline, pyrazolidine, piperidine, piperazine, tetrahydropyridine, tetrahydropyrimidine, tetrahydropyridazine, dihydrofuran, tetrahydrofuran, dihydropyran, tetrahydropyran, dihydrothiophene, tetrahydrothiophene, dihydrothiain (dihydrothiopyran), tetrahydrothiain (tetrahydrothiopyran), dihydrooxazole, tetrahydrooxazole, dihydroisoxazole, tetrahydroisoxazole, dihydrothiazole, tetrahydrothiazole, dihydroisothiazole, tetrahydroisothiazole, morpholine, thiomorpholine, indoline, isoindoline, dihydrobenzofuran, perhydrobenzofuran, dihydroisobenzofuran, perhydroisobenzofuran, dihydrobenzothiophene, perhydrobenzothiophene, dihydroisobenzothiophene, perhydroisobenzothiophene, dihydroindazole, perhydroindazole, dihydroquinoline, tetrahydroquinoline, perhydroquinoline, dihydroisoquinoline, tetrahydroisoquinoline, perhydroisoquinoline, dihydrophthalazine, tetrahydrophthalazine, perhydrophthalazine, dihydronaphthyridine, tetrahydronaphthyridine, perhydronaphthyridine, dihydroquinoxaline, tetrahydroquinoxaline, perhydroquinoxaline, dihydroquinazoline, tetrahydroquinazoline, perhydroquinazoline, dihydrocinnoline, tetrahydrocinnoline, perhydrocinnoline, dihydrobenzoxazole, perhydrobenzoxazole, dihydrobenzothiazole, perhydrobenzothiazole, dihydrobenzimidazole, perhydrobenzimidazole, benzoxazepine, benzoxadiazepine, benzothiazepine, benzothiadiazepine, benzoazepine, benzodiazepine, indolooxazepine, indolotetrahydrooxazepine, indolooxadiazepine, indolotetrahydrooxadiazepine, indolothiazepine, indolotetrahydrothiazepine, indolothiadiazepine, indolotetrahydrothiadiazepine, indoloazepine, indolotetrahydroazepine, indolodiazepine, indolotetrahydrodiazepine, benzofurazan, benzothiadiazole, benzotriazole, camphor, imidazothiazole, dihydrocarbazole, tetrahydrocarbazole, perhydrocarbazole, dihydroacridine, tetrahydroacridine, perhydroacridine, dioxolane, dioxane, dithiolane, dithiane, dioxazine, dithiazine.
Salts
Non-toxic salts of the present invention include all pharmaceutically acceptable salts, for example, general salts, acid addition salts, hydrate salts.
The compounds of formula (I) of the present invention may be converted into the corresponding salts. Non-toxic salts and water-soluble salts are preferred. Suitable salts, for example, include: salts of alkali metals (e.g. potassium, sodium), salts of alkaline earth metals (e.g. calcium, magnesium), ammonium salts, salts of pharmaceutically acceptable organic amines (e.g. tetramethylammonium, triethylamine, methylamine, dimethylamine, cyclopentylamine benzylamine, phenethylamine, piperidine, monoethanolamine, diethanolamine, tris(hydroxymethyl)amine, lysine, arginine, N-methyl-D-glucamine).
The compounds of formula (I) may be converted into the corresponding acid addition salts. Non-toxic acid addition salts and water-soluble acid addition salts are preferred. Suitable salts, for example, include: salts of inorganic acids e.g. hydrochloride, hydrobromide, sulfate, phosphate, nitrate; salts of organic acids e.g. acetate, trifluoroacetate, lactate, tartarate, oxalate, fumarate, maleate, citrate, benzoate, methanesulphonate, ethanesulphonate, benzenesulphonate, toluenesulphonate, isethionate, glucuronate, gluconate.
The compounds of formula (I) and salts thereof may be converted into the corresponding hydrates by conventional means.
In the compounds of the present invention of formula (I), the compounds of the following formula are preferred: 
wherein all the symbols are as hereinbefore defined; 
wherein the symbols are as hereinbefore defined; 
wherein all the symbols are as hereinbefore defined; 
wherein all the symbols are as hereinbefore defined; 
wherein all the symbols are as hereinbefore defined; 
wherein all the symbols are as hereinbefore defined; 
wherein all the symbols are as hereinbefore defined; 
wherein all the symbols are as hereinbefore defined; 
wherein all the symbols are as hereinbefore defined; 
wherein all the symbols are as hereinbefore defined; 
wherein all the symbols are as hereinbefore defined; 
wherein all the symbols are as hereinbefore defined; 
wherein all the symbols are as hereinbefore defined; 
wherein all the symbols are as hereinbefore defined; 
wherein G1 is methyl, halogen atom, nitro or nitrite and the other symbols are as hereinbefore defined; 
wherein all the symbols are as hereinbefore defined.
The compounds in Table1-Table105 and non-toxic salts thereof, and the compounds described in the Examples are more preferred. In the following Tables, Phth is phthalimide, Ph is phenyl, MOM is methoxymethyl, EOM is ethoxymethyl, MEM is (2-methoxyethoxy)methyl, BOM is benzyloxymethyl.
Process for the Preparation
The compounds of the present invention of the formula (I), may be prepared by following methods or the methods described in the Examples.
[1] In the compounds of the present invention of the formula (I), the compound in which R1 is xe2x80x94COOR10, that is the compound of the formula (I-1): 
wherein all the symbols are as hereinbefore defined; may be prepared by following methods (a)-(c).
(a) The compound in which xe2x80x94COOR10 in R1 is not xe2x80x94COOH, and all of R2, R3, R4, R5, R6, R7, R8 and R9 are not xe2x80x94COOH or a group including it, hydroxy or a group including it, amino or a group including it, that is the compound of the formula (I-1a): 
wherein R10-1a is C1-8 alkyl, phenyl, C1-8 alkyl substituted by phenyl or C1-8 alkoxy, oxycarbonyl substituted by phenyl, benzyl or C1-8 alkyl, each of R2-1a R3-1a, R4-1a, R5-1a, R6-1a, R7-1a, R8-1a, R9-1a has the same meaning as R2, R3, R4, R5, R6, R7, R8, R9, with the proviso that, all of R2-1a, R3-1a, R4-1a, R5-1a, R6-1a, R7-1a, R8-1a, R9-1a are not xe2x80x94COOH, hydroxy or amino or groups including them, and the other symbols are as hereinbefore defined;
may be prepared by amidation of the compound of the formula (II): 
wherein all the symbols are as hereinbefore defined;
with the compound of the formula (III): 
wherein all the symbols are as hereinbefore defined.
The method of amidation of the compound of the formula (II) with the compound of the formula (III) is known. It includes the method
(1) via an acyl halide,
(2) via a mixed acid anhydride,
(3) using a condensing agent.
These methods are explained as follows.
(1) The method via an acyl halide, for example, may be carried out in an organic solvent (e.g. chloroform, methylene chloride, diethyl ether or tetrahydrofuran) or without a solvent, using an acyl halide (e.g. oxalyl chloride or thionyl chloride etc.) at xe2x88x9220xc2x0 C. to reflux temperature, and the obtained acyl halide derivative may be reacted with an amine in an organic solvent (e.g. chloroform, methylene chloride, diethyl ether or tetrahydrofuran) in the presence of a tertiary amine (e.g. pyridine, triethyl amine, dimethyl aniline or dimethylaminopyridine) at 0-40xc2x0 C.
(2) The method via a mixed acid anhydride may be carried out, for example, by reacting a carboxylic acid with an acyl halide (e.g. pivaloyl chloride, tosyl chloride or mesyl chloride) or an acid derivative (e.g. ethyl chloroformate or isobutyl chloroformate) in an organic solvent (e.g. chloroform, methylene chloride, diethyl ether or tetrahydrofuran) or without a solvent, in the presence of a tertiary amine (e.g. pyridine, triethylamine, dimethylaniline or dimethylaminopyridine) at 0-40xc2x0 C., and the obtained mixed acid anhydride derivative may be reacted with a corresponding amine in an organic solvent (e.g. chloroform, methylene chloride, diethyl ether or tetrahydrofuran) at 0-40xc2x0 C.
(3) The method using a condensing agent (e.g. 1,3-dicyclohexyl carbodiimide (DCC), 1-ethyl-3-[3-(dimethylamino)propyl]carbodiimide (EDC), 1,1xe2x80x2-carbonyldiimidazole (CDI) or 2-chloro-1-methylpyridinium iodide) may be carried out, for example, by reacting a carboxylic acid with an amine in an organic solvent (e.g. chloroform, methylene chloride, dimethylformamide, diethyl ether or tetrahydrofuran) or without a solvent, optionally in the presence of a tertiary amine (e.g. pyridine, triethylamine, dimethylaniline or dimethylaminopyridine) using a condensing agent, and optionally in the presence of 1-hydroxybenzotriazole (HOBt) at 0-40xc2x0 C.
The reaction described in (1), (2) and (3) may be carried out under an inert gas (e.g. argon, nitrogen) to avoid water in order to obtain a preferable result.
(b) The compound in which xe2x80x94COOR10 in R1 is not xe2x80x94COOH, and at least one of R2, R3, R4, R5, R6, R7, R8 and R9 is xe2x80x94COOH or a group including it, hydroxy or a group including it, amino or a group including it, that is the compound of the formula (I-1b): 
wherein each of R2-1b, R3-1b, R4-1b, R5-1b, R6-1b, R7-1b, R8-1b, R9-1b has the same meaning as R2, R3, R4, R5, R6, R7, R8, R9, with the proviso that, at least one of R2-1b, R3-1b, R4-1b, R5-1b, R6-1b, R7-1b, R8-1b, R9-1b is xe2x80x94COOH, hydroxy or amino or groups including them, and the other symbols are as hereinbefore defined;
may be prepared by deprotection under alkaline conditions, deprotection under acidic conditions, deprotection of a silyl group or hydrogenolysis of the compound having protected xe2x80x94COOH, hydroxy or amino or groups including protected xe2x80x94COOH, hydroxy or amino in the compound of the formula (I-1a), that is the compound of the formula (I-1a1): 
wherein each of R2-1a1, R3-1a1, R4-1a1, R5-1a1, R6-1a1, R7-1a1, R8-1a1, R9-1a1 has the same meaning as R2, R3, R4, R5, R6, R7, R8, R9, with the proviso that, at least one of R2-1a1, R3-1a1, R4-1a1, R5-1a1, R6-1a1, R7-1a1, R8-1a1, R9-1a1 is protected xe2x80x94COOH (e.g. protected by methyl, ethyl, t-butyl and benzyl), protected hydroxy (e.g. protected by methoxymethyl, tetrahydropyranyl, t-butyldimethylsilyl, acetyl, benzyl) or protected amino (e.g. protected by benzyloxycarboyl, t-butoxycarbonyl, trifluoroacetyl) or a group including protected xe2x80x94COOH, hydroxy or amino, and the other symbols are as hereinbefore defined.
Deprotection under alkaline conditions was known, for example, it may be carried out in an organic solvent (e.g. methanol, tetrahydrofuran or dioxane), using an alkali metal hydroxide (e.g. sodium hydroxide, potassium hydroxide or lithium hydroxide), an alkaline earth metal hydroxide (e.g. barium hydroxide or calcium hydroxide) or a carbonate (e.g. sodium carbonate or potassium carbonate), an aqueous solution thereof or mixture thereof at 0-40xc2x0 C.
Deprotection under acidic conditions was known, for example, it may be carried out in an organic solvent (e.g. methylene chloride, chloroform, dioxane, ethyl acetate, anisole), using an organic acid (e.g. acetic acid, trifluoroacetic acid, methansulfonic acid or trimethylsilyl iodide), or an inorganic acid (e.g. hydrochloric acid or sulfuric acid) or a mixture thereof (e.g. hydrogen bromide in acetic acid) at 0-100xc2x0 C.
Deprotection of a silyl group was known, for example, it may be carried out in a water miscible organic solvent (e.g. tetrahydrofuran or acetonitrile), using tetrabutylammonium fluoride at 0-40xc2x0 C.
Hydrogenolysis was known, for example, it may be carried out in a solvent [e.g. ether (such as tetrahydrofuran, dioxane, dimethoxyethane or diethyl ether), alcohol (such as methanol or ethanol), benzene (such as benzene or toluene), ketone (such as acetone or methyl ethyl ketone), nitrile (such as acetonitrile), amide (such as dimethylformamide), water, ethyl acetate, acetic acid or two more mixture thereof], in the presence of a catalyst (e.g. palladium on carbon, palladium black, palladium, palladium hydroxide, platinum dioxide, nickel or Raney-nickel), optionally in the presence of an inorganic acid (e.g. hydrochloric acid, sulfuric acid, hypochlorous acid, boric acid or tetrafluoroboric acid) or an organic acid (e.g. acetic acid, p-toluenesulfonic acid, oxalic acid, trifluoroacetic acid or formic acid), at ordinary or elevated pressure of hydrogen gas or ammonium formate at 0-200xc2x0 C. There is no difficulty in using a salt of acid, when it is carried out using an acid.
(c) The compound in which xe2x80x94COOR10 in R1 is xe2x80x94COOH, that is the compound of the formula (I-1c): 
wherein all the symbols are the same meaning as hereinbefore defined;
may be prepared by deprotection under alkaline conditions, deprotection under acidic conditions or hydrogenolysis of the above compounds of the formula (I-1a) and the formula (I-1b), that is the compound of the formula (I-1ab): 
wherein all the symbols are the same meaning as hereinbefore defined.
The reactions of deprotection under alkaline conditions, deprotection under acidic conditions and hydrogenolysis are known and they may be carried out by the same method as hereinbefore described.
[2] In the compounds of the present invention of the formula (I), the compound in which R1 is xe2x80x94CONHOR10 or xe2x80x94CONHNHR10, that is the compound of the formula (I-2): 
wherein R1-2 is xe2x80x94CONHOR10 or xe2x80x94CONHNHR10, and the other symbols are the same meaning as hereinbefore defined; may be prepared by following methods (a) and (b).
(a) The compound in which R1 is xe2x80x94CONHOR10 or xe2x80x94CONHNHR10, and all of R2, R3, R4, R5, R6, R7, R8 and R9 are not xe2x80x94COOH or a group including it, that is the compound of the formula (I-2a): 
wherein each of R2-2a, R3-2a, R4-2a, R5-2a, R6-2a, R7-2a, R8-2a, R9-2a is the a same meaning as R2, R3, R4, R5, R6, R7, R8 , R9, with the proviso that, all of R2-2a, R3-2a, R4-2a, R5-2a, R6-2a, R7-2a, R8-2a, R9-2a are not xe2x80x94COOH or a group including it, and the other symbols are as hereinbefore defined;
may be prepared by amidation of the compound in which R1 is COOH, and all of R2, R3, R4, R5, R6, R7, R8, R9, are not xe2x80x94COOH or a group including it in the compound of the above formula (I-1), that is the compound of the formula (I-1d): 
wherein all the symbols are the same meaning as hereinbefore defined;
with the compound of the formula (IV):
H2Nxe2x80x94OR10xe2x80x83xe2x80x83(IV)
wherein R10 is the same meaning as hereinbefore defined; or the compound of the formula (V):
xe2x80x83H2Nxe2x80x94NHR10xe2x80x83xe2x80x83(V)
wherein R10 is the same meaning as hereinbefore defined;
if necessary, followed by deprotection under alkaline conditions and/or deprotection under acidic conditions and/or hydrogenolysis.
This reaction of amidation, the reactions of deprotection under alkaline conditions, deprotection under acidic conditions and hydrogenolysis are known, and may be carried out by the same method as hereinbefore described.
(b) The compound in which R1 is xe2x80x94CONHOR10 or xe2x80x94CONHNHR10, and at least one of R2, R3, R4, R5, R6, R7, R8 and R9 is xe2x80x94COOH or a group including it, that is the compound of the formula (I-2b): 
wherein each of R2-2b, R3-2b, R4-2b, R5-2b, R6-2b, R7-2b, R8-2b, R9-2b is the a same meaning as R2, R3, R4, R5, R6, R7, R8, R9, with the proviso that, at least one of R2-2b, R3-2b, R4-2b, R5-2b, R6-2b, R7-2b, R8-2b, R9-2b is xe2x80x94COOH or a group including it, and the other symbols are as hereinbefore defined;
may be prepared by deprotection under alkaline conditions, deprotection under acidic conditions or hydrogenolysis of the compound having protected xe2x80x94COOH or a group including protected xe2x80x94COOH in the compound of the above formula (I-2a), that is the compound of the formula (I-2a1): 
wherein each of R2-2a1, R3-2a1, R4-2a1, R5-2a1, R6-2a1, R7-2a1, R8-2a1, R9-2a1 has the same meaning as R2, R3, R4, R5, R6, R7, R8, R9, with the proviso that, at least one of R2-2a1, R3-2a1, R4-2a1, R5-2a1, R6-2a1, R7-2a1, R8-2a1, R9-2a1, is protected xe2x80x94COOH (e.g. protected by methyl, ethyl, t-butyl and benzyl) or a group including protected xe2x80x94COOH, and the other symbols are as hereinbefore defined.
The reactions of deprotection under alkaline conditions, deprotection under acidic conditions and hydrogenolysis are known, and may be carried out by the same method as hereinbefore described.
[3] In the compounds of the present invention of the formula (I), the compound in which R1 is xe2x80x94(CH2)nSR50, that is the compound of the formula (I-3): 
wherein R1-3 is xe2x80x94(CH2)nSR50, and the other symbols are as hereinbefore defined; may be prepared by following methods (a) and (b).
(a) The compound in which R1 is xe2x80x94(CH2)nSR50, and all of R2, R3, R4, R5, R6, R7, R8 and R9 are not xe2x80x94COOH or a group including it, that is the compound of the formula (I-3a): 
wherein each of R2-3a, R3-3a, R4-3a, R5-3a, R6-3a, R7-3a, R8-3a, R9-3a has the same meaning as R2, R3, R4, R5, R6, R7, R8, R9, with the proviso that, all of R2-3a, R3-3a, R4-3a, R5-3a, R6-3a, R7-3a, R8-3a, R9-3a are not xe2x80x94COOH or a group including it, and the other symbols are as hereinbefore defined;
may be prepared by reaction of the compound of the formula (VI): 
wherein X is halogen atom and the other symbols are as hereinbefore defined; and the compound of the formula (VII):
R501SKxe2x80x83xe2x80x83(VII)
wherein R501 is C1-8 alkyl, xe2x80x94COR52 or xe2x80x94SR531, in which R531 is C1-8 alkyl or phenyl.
The compound in which R50 is hydrogen or xe2x80x94SH may be prepared by a reaction of deprotection of the compound obtained by the above method.
The above method was known, for example, it may be carried out by refluxing in an organic solvent (e.g. acetone, tetrahydrofuran).
The continuous reaction of deprotection was known, for example, it may be carried out in an organic solvent (e.g. methanol, tetrahydrofuran or dioxane), using an alkali metal hydroxide (e.g. sodium hydroxide, potassium hydroxide or lithium hydroxide), an alkaline earth metal hydroxide (e.g. barium hydroxide or calcium hydroxide) or a carbonate (e.g. sodium carbonate or potassium carbonate), an aqueous solution thereof or mixture thereof at 0-40xc2x0 C.
(b) The compound in which R1 is xe2x80x94(CH2)nSR50, and at least one of R2, R3, R4, R5, R6, R7, R8 and R9 is xe2x80x94COOH or a group including it, that is the compound of the formula (I-3b): 
wherein each of R2-3b, R3-3b, R4-3b, R5-3b, R6-3b, R7-3b, R8-3b, R9-3b has the same meaning as R2, R3, R4, R5, R6, R7, R8, R9, with the proviso that, at least one of R2-3b, R3-3b, R4-3b, R5-3b, R6-3b, R7-3b, R8-3b, R9-3b is xe2x80x94COOH or a group including it, and the other symbols are as hereinbefore defined;
may be prepared by deprotection under alkaline conditions, deprotection under acidic conditions or hydrogenolysis of the compound having protected xe2x80x94COOH or a group including protected xe2x80x94COOH in the compound of the above formula (I-3a), that is the compound of the formula (I-3a1): 
wherein each of R2-3a1, R3-3a1, R4-3a1, R5-3a1, R6-3a1, R7-3a1, R8-3a1, R9-3a1 has the same meaning as R2, R3, R4, R5, R6, R7, R8, R9, with the proviso that, at least one of R2-3a1, R3-3a1, R4-3a1, R5-3a1, R6-3a1, R7-3a1, R8-3a1, R9-3a1 is protected xe2x80x94COOH (e.g. protected by methyl, ethyl, t-butyl and benzyl) or a group including protected xe2x80x94COOH, and the other symbols are as hereinbefore defined.
The reactions of deprotection under alkaline conditions, deprotection under acidic conditions and hydrogenolysis are known, and may be carried out by the same method as hereinbefore described.
[4] In the compounds of the present invention of the formula (I), the compound in which R1 is xe2x80x94Yxe2x80x94PO(OR51)2, that is the compound of the formula (I-4): 
wherein R1-4 is xe2x80x94Yxe2x80x94PO(OR51)2, and the other symbols are as hereinbefore defined; may be prepared by following methods (a)-(d).
(a) The compound in which R1 is xe2x80x94Yxe2x80x94PO(OR51)2, in which Y1 is xe2x80x94Oxe2x80x94 and the other symbols are as hereinbefore defined; and all of R2, R3, R4, R5, R6, R7, R8 and R9 are not xe2x80x94COOH or a group including it, that is the compound of the formula (I-4a): 
wherein each of R2-4a, R3-4a, R4-4a, R5-4a, R6-4a, R7-4a, R8-4a, R9-4a has the same meaning as R2, R3, R4, R5, R6, R7, R8, R9, with the proviso that, all of R2-4a, R3-4a, R4-34a, R5-4a, R6-4a, R7-4a, R8-4a, R9-4a are not xe2x80x94COOH or a group including it, and the other symbols are as hereinbefore defined;
may be prepared by reaction of the compound of the formula (VIII): 
wherein all the symbols are as hereinbefore defined; and the compound of the formula (IX): 
wherein R511 is C1-8 alkyl, phenyl or a known protecting group of phosphoric acid and the other symbols are as hereinbefore defined. Furthermore, the compound of the formula (I-4a) may be prepared, followed by deprotection of the compound having a protecting phosphoric acid.
The above reaction was known, for example, it may be carried out in an organic solvent (e.g. pyridine) at 0-40xc2x0 C.
The reaction of deprotection of phosphoric acid was known, for example, it may be carried out in an organic solvent (e.g. pyridine), using zinc acetate at 0-40xc2x0 C.
(b) The compound in which R1 is xe2x80x94Y1xe2x80x94PO(OR51)2, and at least one of R2, R3, R4, R5, R6, R7, R8 and R9 is xe2x80x94COOH or a group including it, that is the compound of the formula (I-4b): 
wherein each of R2-4b, R3-4b, R4-4b, R5-4b, R6-4b, R7-4b, R8-4b, R9-4b has the same meaning as R2, R3, R4, R5, R6, R7, R8, R9, with the proviso that, at least one of R2-4b, R3-4b, R4-4b, R5-4b, R6-4b, R7-4b, R8-4b, R9-4b is xe2x80x94COOH or a group including it, and the other symbols are as hereinbefore defined;
may be prepared by deprotection under alkaline conditions, deprotection under acidic conditions or hydrogenolysis of the compound having protected xe2x80x94COOH or a group including protected xe2x80x94COOH in the compound of the above formula (I-4a), that is the compound of the formula (I-4a1); 
wherein each of R2-4a1, R3-4a1, R4-4a1, R5-4a1, R6-4a1, R7-4a1, R8-4al, R9-4a1 has the same meaning as R2, R3, R4, R5, R6, R7, R8, R9, with the proviso that, at least one of R2-4a1, R3-4a1, R4-4a1, R5-4a1, R6-4a1, R7-4a1, R8-4a1, R9-4a1 is protected xe2x80x94COOH (e.g. protected by methyl, ethyl, t-butyl and benzyl) or a group including protected xe2x80x94COOH, and the other symbols are as hereinbefore defined.
The reactions of deprotection under alkaline conditions, deprotection under acidic conditions and hydrogenolysis are known, and may be carried out by the same method as hereinbefore described.
(c) The compound in which R1 is xe2x80x94Y2xe2x80x94PO(OR51)2, in which Y2 is a single bond or xe2x80x94CH2xe2x80x94 and the other symbols are as hereinbefore defined; and all of R2, R3, R4, R5, R6, R7, R8 and R9 are not xe2x80x94COOH or a group including it, that is the compound of the formula (I-4c): 
wherein each of R2-4c, R3-4c, R4-4c, R5-4c, R6-4c, R7-4c, R8-4c, R9-4c has the same meaning as R2, R3, R4, R5, R6, R7, R8, R9, with the proviso that, all of R2-4c, R3-4c, R4-4c, R5-4c, R6-4c, R7-4c, R8-4c, R9-4c are not xe2x80x94COOH or a group including it, and the other symbols are as hereinbefore defined;
may be prepared by reaction of the compound of the formula (X): 
wherein all the symbols are as hereinbefore defined; and the compound of the formula (XI) or (XII):
(R511O)3Pxe2x80x83xe2x80x83(XI)
or
(R511O)2POKxe2x80x83xe2x80x83(XII)
wherein all the symbols are as hereinbefore defined. Furthermore, the compound of the formula (I-4c) may be prepared, followed by deprotection of the compound having a protecting phosphoric acid.
The above reaction was known, for example, it may be carried out in an organic solvent (e.g. tetrahydrofuran, dimethylformamide) at 0-120xc2x0 C.
The reaction of deprotection of phosphoric acid was known, and it may be carried out by the same method as hereinbefore described.
(d) The compound in which R1 is xe2x80x94Y2xe2x80x94PO(OR51)2, and at least one of R2, R3, R4, R5, R6, R7, R8 and R9 is xe2x80x94COOH or a group including it, that is the compound of the formula (I-4d): 
wherein each of R2-4d, R3-4d, R4-4d, R5-4d, R6-4d, R7-4d, R8-4d, R9-4d has the same meaning as R2, R3, R4, R5, R6, R7, R8, R9, with the proviso that, at least one of R2-4d, R3-4d, R4-4d, R5-4d, R6-4d, R7-4d, R8-4d, R9-4d is xe2x80x94COOH or a group including it, and the other symbols are as hereinbefore defined;
may be prepared by deprotection under alkaline conditions, deprotection under acidic conditions or hydrogenolysis of the compound having protected xe2x80x94COOH or a group including protected xe2x80x94COOH in the compound of the above formula (I-4c), that is the compound of the formula (I-4c1): 
wherein each of R2-4c1, R3-4c1, R4-4c1, R5-4c1, R6-4c1, R7-4c1, R8-4c1, R9-4c1 has the same meaning as R2, R3, R4, R5, R6, R7, R8, R9, with the proviso that, at least one of R2-4c1, R3-4c1, R4-4c1, R5-4c1, R6-4c1, R7-4c1, R8-4c1, R9-4c1 is protected xe2x80x94COOH (e.g. protected by methyl, ethyl, t-butyl and benzyl) or a group including protected xe2x80x94COOH, and the other symbols are as hereinbefore defined.
The reactions of deprotection under alkaline conditions, deprotection under acidic conditions and hydrogenolysis are known, and may be carried out by the same method as hereinbefore described.
The reactions of deprotection in the present invention are common reactions of deprotection as will be apparent to those skilled in the art, for example, deprotection under alkaline conditions, deprotection under acidic conditions or hydrogenolysis. The desired compound of the present invention may be easily prepared using these protecting groups.
As will be apparent to those skilled in the art, methyl, ethyl, t-butyl or benzyl may be used as protecting groups for carboxyl, but other groups which may be removed easily and selectively are also preferred. For example, the groups described in T. W. Greene, Protective Groups in Organic Synthesis, Wiley, New York, 1991, may be used.
Methoxymethyl, tetrahydropyranyl, t-butyldimethylsilyl, acetyl or benzyl may be used as protecting groups for hydroxy, but other groups which may be removed easily and selectively are also preferred. For example, the groups described in T. W. Greene, Protective Groups in Organic Synthesis, Wiley, New York, 1991, may be used.
Benzyloxycarbonyl, t-butoxycarbonyl or trifluoroacetyl maybe used as protecting groups for amino, but other groups which may be removed easily and selectively are also preferred. For example, the groups described in T. W. Greene, Protective Groups in Organic Synthesis, Wiley, New York, 1991, may be used.
The compounds of the formulae (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), (X), (XI), and (XII) are known per se or may be prepared by known methods.
In each reaction in the present specification, products may be purified by conventional techniques. For example, purification may be carried out by distillation at atmospheric or reduced pressure, by high performance liquid chromatography, by thin layer chromatography or by column chromatography using silica gel or magnesium silicate, by washing or by recrystallization. Purification may be carried out after each reaction, or after a series of reactions.
Pharmacological Activities
The potency of inhibitory activity against each matrix metalloproteinase of the compound of the formula (I) was confirmed as below.
(1) Inhibitory Activity Against Gelatinase A
[Method]
The progelatinase A (5 xcexcl; in assay buffer (40 xcexcl)) was purified from human normal skin dermal fibroblasts (HNDF). It was activated by the addition of 10 mM of p-aminophenylmercuric acetate (APMA) (5 xcexcg) for 1 hour at 37xc2x0 C.
A mixture of the synthetic substrate (MOCAc-Pro-Leu-Gly-A2pr(Dnp)-Ala-Arg-NH2) (130 xcexcl; a final concentration 13.5 xcexcM) and a solution (20 xcexcl) with or without various concentrations of the test compound was preincubated for 5 minutes at 37xc2x0 C.
The solution of activated gelatinase A (50 xcexcl/well) was mixed with the mixture and the mixture was incubated for 15 minutes at 37xc2x0 C. The enzyme reaction was started. The enzyme activity was represented by increasing value of fluorescent intensity [Ex=325 nm (Ex)/393 nm (Em)] per 1 minute. Inhibitory activity was represented by inhibitory percentage (%) per enzyme activity without the test compound. For example, IC50 of the compound as Example 71 is 0.50 nM.
(2) Inhibitory Activity Against Collagenase
[Method]
The procollagenase (5 xcexcl; in assay buffer (105 xcexcl)) was purified from human normal skin dermal fibroblasts (HNDF). It was activated by the addition of 1 mg/ml Trypsin (45 xcexcl) for 1 minute at 37xc2x0 C. Trypsin was inactivated by addition of 5 mg/ml soybean trypsin inhibitor (SBTI; 50 xcexcl).
A mixture of the synthetic substrate (Ac-Pro-Leu-Gly-[2-mercapto-4-methyl-pentanoyl]-Leu-Gly-OEt) (105 xcexcl; a final concentration 1.33 mM) and a solution (20 xcexcl) with or without various concentrations of the test compound was preincubated for 5 minutes at 26xc2x0 C.
The solution of activated enzyme (75 xcexcl/tube, 50 xcexcl) was mixed with the mixture and the mixture was incubated for 10 minutes at 26xc2x0 C.
Absorption at 324 nm was measured at 40 points over 10 minutes. Vmax value was determined as measured value in 30 points therein. For example, IC50 of the compound as Example 71 is 2.5 xcexcM.
(3) Inhibitory Activity Against Stromelysin ps [Method]
The mixture of human stromelysin (Yagai; 9 volume) and 10 mM p-aminophenylmercury acetate (1 volume) was activated for 20 hours at 37xc2x0 C. A solution of the test compound in dimethylsulfoxide (10 xcexcl) and 0.5 mM solution (10 xcexcl) of 10 mM solution of the synthetic substrate NFF-3 (Mca-Arg-Pro-Lys-Pro-Val-Glu-Nva-Trp-Arg-Lys(DNP)-NH2., Nva: norvaline, Peptide Laboratory) in dimethylsulfoxide diluted by water were added to assay buffer (50 mM tris-HCl, 10 mM CaCl2, 0.05% Brij35, 0.02% NaN3 (pH 7.5)) (150 xcexcl). Furthermore, assay buffer (30 xcexcl) was added to the mixture. The mixture was incubated for 10 minutes at 37xc2x0 C. The reaction was started by addition of a solution of the above activated stromelysin solution (50 xcexcl). The enzyme activity was represented by increasing value of fluorescent intensity [Ex=325 nm (Ex)/393 nm (Em)] per 1 minute. Inhibitory activity was represented by inhibitory percentage (%) per enzyme activity without the test compound. For example, IC50 of the compound as Example 71 is 26 nM.
Toxicity
The toxicity of the compounds of the present invention is very low and therefore, the compounds may be considered safe for pharmaceutical use.
Application for Pharmaceuticals
Inhibition of matrix metalloproteinase, for example, gelatinase, stromelysin or collagenase, is useful for prevention and/or treatment of diseases, for example, rheumatoid diseases, arthrosteitis, unusual bone resorption, osteoporosis, periodontitis, interstitial nephritis, arteriosclerosis, pulmonary emphysema, cirrhosis, cornea injury, metastasis of, invasion of or growth of tumor cells, autoimmune disease (e.g. Crohn""s disease, Sjogren""s syndrome), disease caused by vascular emigration or infiltration of leukocytes, arterialization, multiple sclerosis, aorta aneurysm, endometriosis in animals including human beings, especially human beings.
For the purpose above described, the compounds of formulae (I) of the present invention and non-toxic salts, acid addition salts or hydrates may be normally administered systemically or locally, usually by oral or parenteral administration.
The doses to be administered are determined depending upon, for example, age, body weight, symptom, the desired therapeutic effect, the route of administration, and the duration of the treatment. In the human adult, the doses per person are generally from 1 mg to 1000 mg, by oral administration, up to several times per day, and from 1 mg to 100 mg, by parenteral administration (preferably intravenous administration), up to several times per day, or continuous administration from 1 to 24 hours per day from vein.
As mentioned above, the doses to be used depend upon various conditions. Therefore, there are cases in which doses lower than or greater than the ranges specified above may be used.
The compounds of the present invention may be administered in the form of, for example, solid compositions, liquid compositions or other compositions for oral administration, injections, liniments or suppositories for parenteral administration.
Solid compositions for oral administration include compressed tablets, pills, capsules, dispersible powders, and granules.
Capsules include hard capsules and soft capsules.
In such compositions, one or more of the active compound(s) may be admixed with at least one inert diluent (such as lactose, mannitol, glucose, hydroxypropyl cellulose, microcrystalline cellulose, starch, polyvinylpyrrolidone or magnesium metasilicate aluminate). The compositions may also comprise, as is normal practice, additional substances other than inert diluents: e.g. lubricating agents (such as magnesium stearate), disintegrating agents (such as cellulose calcium glycolate), stabilizing agents, and agents to assist dissolution (such as glutamic acid or aspartic acid).
The tablets or pills may, if desired, be coated with a film of gastric or enteric material (such as sugar, gelatin, hydroxypropyl cellulose or hydroxypropylmethyl cellulose phthalate), or be coated with two or more films. And further, coating may include containment within capsules of absorbable materials such as gelatin.
Liquid compositions for oral administration include pharmaceutically acceptable emulsions, solutions, syrups and elixirs. In such compositions, one or more of the active compound(s) may be contained in an inert diluent(s) commonly used in the art (e.g. purified water or ethanol). Besides inert diluents, such compositions may also comprise adjuvants (such as wetting agents or suspending agents), sweetening agents, flavouring agents, perfuming agents, and preserving agents.
Other compositions for oral administration include spray compositions which may be prepared by known methods and which comprise one or more of the active compound(s). Spray compositions may comprise additional substances other than inert diluents: e.g. stabilizing agents (such as sodium sulfate), isotonic buffer (such as sodium chloride, sodium citrate or citric acid). For preparation of such spray compositions, for example, the method described in the U.S. Pat. No. 2,868,691 or U.S. Pat. No. 3,095,355 may be used.
Injections for parenteral administration include sterile aqueous or non-aqueous solutions, suspensions and emulsions. Aqueous solutions and suspensions may include distilled water for injection or physiological salt solution. Non-aqueous solutions and suspensions may include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, alcohol such as ethanol or POLYSORBATE80 (registered trade mark).
Injections may comprise additional ingredients other than inert diluents: e.g. preserving agents, wetting agents, emulsifying agents, dispersing agents, stabilizing agents, assisting agents such as agents to assist dissolution (e.g. glutamic acid or aspartic acid).
They may be sterilized for example, by filtration through a bacteria-retaining filter, by incorporation of sterilizing agents in the compositions or by irradiation. They may also be manufactured in the form of sterile solid compositions which may be dissolved in sterile water or some other sterile diluent(s) for injection immediately before use.
Other compositions for parenteral administration include liquids for external use, and endermic liniments, ointment, suppositories for rectal administration and pessaries for vaginal administration which comprise one or more of the active compound(s) and may be prepared by methods known per se.